Role of AP-1 complex in angiotensin II-mediated transforming growth factor-beta expression and growth of smooth muscle cells: using decoy approach against AP-1 binding site.
The cis element double-stranded oligodeoxynucleotides (ODN) ("decoy") approach has enabled us to clarify the responsible elements. Using this approach, we transfected AP-1 decoy ODN into VSMC cultured from WKY rats (WKY Adu) which produce latent TGF-beta, but not active TGF-beta, and Sprague-Dawley rats (CNC) which produce active and latent TGF-beta under Ang II-stimulation. AP-1, but not mismatched, decoy ODN abolished Ang II-stimulated TGF-beta gene expression and production in both Adu and CNC. However, AP-1 decoy did not alter DNA, RNA, and protein synthesis in Adu. In contrast, cell number was increased under Ang II stimulation by AP-1 decoy ODN in CNC without significant change in RNA and protein synthesis. These results showed that Ang II stimulated TGF-beta production through the AP-1 complex. In VSMC that produce active TGF-beta (CNC), the AP-1 complex stimulated by Ang II may inhibit cell growth through active TGF-beta production. Overall, this study demonstrates the utility of the decoy approach to study the specific function of cis elements in endogenous gene regulation such as Ang II-induced TGF-beta expression.
Morishita
R
R
Division of Cardiovascular Medicine, Brigham and Women's Hospital, Harvard Medical School, Boston, Massachusetts, USA.
Gibbons
G H
GH
Horiuchi
M
M
Kaneda
Y
Y
Ogihara
T
T
Dzau
V J
VJ
D000804:Angiotensin II / Q000494:pharmacology*; D000818:Animals; D002452:Cell Count / Q000187:drug effects; D002478:Cells, Cultured; D016222:Fibroblast Growth Factor 2 / Q000494:pharmacology; D005786:Gene Expression Regulation / Q000187:drug effects*; D007930:Leucine / Q000378:metabolism; D024510:Muscle Development; D009131:Muscle, Smooth, Vascular / Q000187:drug effects / Q000254:growth & development*; D009838:Oligodeoxyribonucleotides / Q000494:pharmacology; D051381:Rats; D011919:Rats, Inbred Strains; D018808:Transcription Factor AP-1 / Q000502:physiology*; D014162:Transfection / Q000235:genetics; D016212:Transforming Growth Factor beta / Q000378:metabolism* / Q000502:physiology; D014529:Uridine / Q000378:metabolism
D009838:Oligodeoxyribonucleotides; D018808:Transcription Factor AP-1; D016212:Transforming Growth Factor beta; D016222:Fibroblast Growth Factor 2; D000804:Angiotensin II; D007930:Leucine; D014529:Uridine